I'm thinking of making an experiment of storing purpurea inoculation material for a year or more using
various methods of storage.
For now I got some sclerotia getting the "cold treatment".
Then I'm thinking of putting sclerotia on agar media and incubate at 25C so it grows stroma and later collect
spores.
For collecting spores - puncture shroom heads with a needle and collect spores on a piece of glass.
After this go two parallel ways - store spores and store honeydew.
For storing spores :
Put collected spores in small amount of distilled water and using arterial blood collecting system
with vacuumtubes suck it into storage tube through a needle.
For honeydew :
Germinate spores on agar medium(thinking of MS basal media with added dextrose or will go with potato-dextose-agar)
and when the honeydew stage is reached, collect it using the same method as above.
Storage format :
some tubes will be put in a fridge at -4C,
some tubes will be put at +1..+4,
some tubes in liquid nitrogen.
Problematic points I can think of for now :
* I'm unsure about storing spores in water. Will it damage them or not.
Maybe there is some better suited liquid medium to store them in.
* If honeydew should be better stored as is or should it too be made into suspension.
* will deep freezing in nitrogen rupture the cells of biomaterial.
After certain time passes, I will try to inoculate media using stored material and see how it goes.
I'm writing this to induce discussion/critique/tips/etc.
Maybe I'm completely wrong on everything, have read so many papers/literature/posts during the last days about claviceps, that
it's all big chaos in my brain and needs ordering.
various methods of storage.
For now I got some sclerotia getting the "cold treatment".
Then I'm thinking of putting sclerotia on agar media and incubate at 25C so it grows stroma and later collect
spores.
For collecting spores - puncture shroom heads with a needle and collect spores on a piece of glass.
After this go two parallel ways - store spores and store honeydew.
For storing spores :
Put collected spores in small amount of distilled water and using arterial blood collecting system
with vacuumtubes suck it into storage tube through a needle.
For honeydew :
Germinate spores on agar medium(thinking of MS basal media with added dextrose or will go with potato-dextose-agar)
and when the honeydew stage is reached, collect it using the same method as above.
Storage format :
some tubes will be put in a fridge at -4C,
some tubes will be put at +1..+4,
some tubes in liquid nitrogen.
Problematic points I can think of for now :
* I'm unsure about storing spores in water. Will it damage them or not.
Maybe there is some better suited liquid medium to store them in.
* If honeydew should be better stored as is or should it too be made into suspension.
* will deep freezing in nitrogen rupture the cells of biomaterial.
After certain time passes, I will try to inoculate media using stored material and see how it goes.
I'm writing this to induce discussion/critique/tips/etc.
Maybe I'm completely wrong on everything, have read so many papers/literature/posts during the last days about claviceps, that
it's all big chaos in my brain and needs ordering.